Spatiotemporal Imaging Research Articles

Vision-Based Prediction of Flashover Using Transformers and Convolutional Long Short-Term Memory Model

The prediction of fire growth is crucial for effective firefighting and rescue operations. Recent advancements in vision-based techniques using RGB vision and infrared (IR) thermal imaging data, coupled with artificial intelligence and deep learning techniques, have shown promising solutions to be applied in the detection of fire and the prediction of its behavior. This study introduces the use of Convolutional Long Short-term Memory (ConvLSTM) network models for predicting room fire growth by analyzing spatiotemporal IR thermal imaging data acquired from full-scale room fire tests. Our findings revealed that SwinLSTM, an enhanced version of ConvLSTM combined with transformers (a deep learning architecture based on a new mechanism called multi-head attention) for computer vision purposes, can be used for the prediction of room fire flashover occurrence. Notably, transformer-based ConvLSTM deep learning models, such as SwinLSTM, demonstrate superior prediction capability, which suggests a new vision-based smart solution for future fire growth prediction tasks. The main focus of this work is to perform a feasibility study on the use of a pure vision-based deep learning model for analysis of future video data to anticipate behavior of fire growth in room fire incidents.

Decoding the interplay between m6A modification and stress granule stability by live-cell imaging.

N6-methyladenosine (m6A)-modified mRNAs and their cytoplasmic reader YTHDFs are colocalized with stress granules (SGs) under stress conditions, but the interplay between m6A modification and SG stability remains unclear. Here, we presented a spatiotemporal m6A imaging system (SMIS) that can monitor the m6A modification and the translation of mRNAs with high specificity and sensitivity in a single live cell. SMIS showed that m6A-modified reporter mRNAs dynamically enriched into SGs under arsenite stress and gradually partitioned into the cytosol as SG disassembled. SMIS revealed that knockdown of YTHDF2 contributed to SG disassembly, resulting in the fast redistribution of mRNAs from SGs and rapid recovery of stalled translation. The mechanism is that YTHDF2 can regulate SG stability through the interaction with G3BP1 in m6A-modified RNA-dependent manner. Our results suggest a mechanism for the interplay between m6A modification and SG through YTHDF2 regulation.

Remote detection of polluted gases based on thermal infrared spectral imaging in the field

This paper reports the new progress of long wave infrared remote sensing in the field, focusing on the implementation process of window scanning spatiotemporal modulation Fourier spectroscopic imaging technology. Utilizing the self-developed CHIPED-1 device, the spatial modulation interference was achieved through the use of a cone mirror Michelson interferometer. By combining it with a cooled long-wave infrared focal plane detector component and applying data processing steps such as acquisition, recombination, calibration, etc., high-spectral resolution imaging in long-wave infrared region was accomplished. The detection sensitivity index nesr (Noise Equivalent Spectral Radiance) of the self-developed CHIPED-1 long wave infrared hyperspectral imaging principle experimental device reaches 5.6 × 10−8w/(cm−1.sr.cm2) in single pixel，Equivalent to commercial time modulation interferometric hyperspectral imagers; It reflects the progressiveness of the technology, and leaves much room for improvement.By testing the transmittance curve of polypropylene film, the spectral response range of CHIPED-1 infrared hyperspectral imaging principle experimental device reached 11.5 μm.The article also studied the hyperspectral imaging detection method for two-dimensional distributed chemical gas VOCs, taking the detection experiments of field high-rise buildings and ether gas as examples.Under complex backgrounds and low experimental concentrations, the presence of ether vapor cannot be observed from the infrared spectrum slices at the same wave number. However, after differential spectral processing, the spatial distribution of ether vapor can be clearly seen.The hyperspectral method is applied in the field of infrared detection of organic vapor VOCs, which has many advantages over wide-band thermal imaging methods, such as high sensitivity, strong anti-interference ability, and wide recognition range.

Asynchronous wide-field transient absorption microscopy.

We have developed a new, to the best of our knowledge, femtosecond laser spatiotemporal imaging technique, named asynchronous wide-field transient absorption microscopy (AWTAM), which does not require phase synchronization between an optical chopper and a complementary metal-oxide-semiconductor (CMOS) camera. By presenting a theoretical scheme, an image reconstruction algorithm, and experimentally imaging the photocarrier diffusion process in a 2D layered semiconductor, our technique has been comprehensively demonstrated. Our technique is widely applicable for fundamental photo-physical research and industrial applications, with the advantages of lower hardware cost as well as higher imaging speed.

Imaging nitric oxide dynamics in endoplasmic reticulum stress with a tailor-made near-infrared fluorescence probe

Spatiotemporal fluorescence imaging of NO dynamics during endoplasmic reticulum (ER) stress is crucial for revealing the molecular mechanisms behind NO-mediated neurological and vascular diseases. However, visualizing in vivo NO dynamics during ER stress through fluorescence imaging is challenging due to interference from light scattering and tissue autofluorescence, which limit imaging penetration and reduce the signal-to-noise ratio. Herein, we report the design of a near-infrared fluorescence probe (ER-NO705) for imaging NO dynamics in ER stress. The probe showed an ultrafast and highly selective response manner towards NO, distinguishing it from other biologically relevant species. Notably, ER-NO705 exhibited excellent ER target-ability, which enabled it could be used for spatiotemporal imaging of NO dynamics in ER stress both in living cells, zebrafish and mice. Overall, this study offers a practical imaging agent for studying the role played by NO in ER stress and offering insights into the biological roles of NO in the ER.

The GTP-tubulin cap is not the determinant of microtubule end stability in cells.

Microtubules are dynamic cytoskeletal polymers essential for cell division, motility, and intracellular transport. Microtubule dynamics are characterized by dynamic instability-the ability of individual microtubules to switch between phases of growth and shrinkage. Dynamic instability can be explained by the GTP-cap model, suggesting that a "cap" of GTP-tubulin subunits at the growing microtubule end has a stabilizing effect, protecting against microtubule catastrophe-the switch from growth to shrinkage. Although the GTP-cap is thought to protect the growing microtubule end, whether the GTP-cap size affects microtubule stability in cells is not known. Notably, microtubule end-binding proteins, EBs, recognize the nucleotide state of tubulin and display comet-like localization at growing microtubule ends, which can be used as a proxy for the GTP-cap. Here, we employ high spatiotemporal resolution imaging to compare the relationship between EB comet size and microtubule dynamics in interphase LLC-PK1 cells to that measured in vitro. Our data reveal that the GTP-cap size in cells scales with the microtubule growth rate in the same way as in vitro. However, we find that microtubule ends in cells can withstand transition to catastrophe even after the EB comet is lost. Thus, our findings suggest that the presence of the GTP-cap is not the determinant of microtubule end stability in cells.

On the Origin of a Broad Quasiperiodic Fast-propagating Wave Train: Unwinding Jet as the Driver

Large-scale extreme-ultraviolet waves commonly exhibit as single wave front and are believed to be caused by coronal mass ejections. Utilizing high spatiotemporal resolution imaging observations from the Solar Dynamics Observatory, we present two sequentially generated wave trains originating from the same active region: a narrow quasiperiodic fast-propagating (QFP) wave train that propagates along the coronal loop system above the jet and a broad QFP wave train that travels along the solar surface beneath the jet. The measurements indicate that the narrow QFP wave train and the accompanying flare’s quasiperiodic pulsations (QPPs) have nearly identical onsets and periods. This result suggests that the accompanying flare process excites the observed narrow QFP wave train. However, the broad QFP wave train starts approximately 2 minutes before the QPPs of the flare, but it is consistent with the interaction between the unwinding jet and the solar surface. Moreover, we find that the period of the broad QFP wave train, approximately 130 s, closely matches that of the unwinding jet. This period is significantly longer than the 30 s period of the accompanying flare’s QPPs. Based on these findings, we propose that the intermittent energy release of the accompanying flare excited the narrow QFP wave train confined propagating in the coronal loop system. The unwinding jet, rather than the intermittent energy release in the accompanying flare, triggered the broad QFP wave train propagating along the solar surface.

Abstract C016: Optical Techniques to Democratize Intravital Quantification of Metabolic Heterogeneity

Abstract Since metabolic and vascular adaptations are essential for tumor cell survival, a technique capable of tracking metabolic heterogeneity at both the bulk tumor and cellular scales over extended timelines is necessary. To address this technological need, we have developed a multi-scale optical microscope relying on exogenous fluorescent reporters for dynamic spatiotemporal imaging of major metabolic pathways in vivo. This microscope utilizes a low-cost CMOS detector and LEDs for excitation to enable fluorescence microscopy in resource-limited settings. Here, we implement our system to assess changes in bulk tumor and cellular metabolic heterogeneity as vasculature is perturbed. 4T1 breast tumor-bearing mice were treated with combretastatin A-1 (a vascular disruptor), implanted with window chambers, and temporally imaged (n=5/group). Fluorescence imaging and Gabor filter/Djikstra segmentation approaches enabled metabolic and vascular analyses, respectively. We demonstrated the system’s ability to acquire co-registered images of vasculature, morphology, and distinct metabolic endpoints reporting on glucose uptake, fatty acid uptake, and mitochondrial metabolism at both wide-field and high resolutions. At 12 hours post-treatment, the tumors’ metabolism shifted from mitochondrial respiration to glycolysis in avascular regions. Further, we report a higher average vascular density in untreated versus treated mice at 108 hours post-treatment (p<0.05, Kolmogorov-Smirnov test). On exploring the relationship between metabolic activity and vasculature, regions with high local tortuosity in the untreated group exhibited a higher reliance on fatty acid uptake than high tortuosity regions in the treatment group, indicating these cells’ reliance on fatty acids within a complex vascular network. Our multiplexed imaging strategy enables the quantification of tumor metabolism and vasculature over multiple length scales, which could be leveraged to study adaptations following clinical therapies. Citation Format: Enakshi Sunassee. Optical Techniques to Democratize Intravital Quantification of Metabolic Heterogeneity [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C016.

Sensitive Cancer Hypoxia Detection via a Dual-Locking Fluorescence Response System Using Two Hypoxia Indicators.

Hypoxia is intricately associated with various diseases, including ischemia, vascular disorders, and cancer. Particularly in cancer cells, hypoxia promotes tumor growth, cell proliferation, migration, and invasion and enhances treatment resistance, making its detection crucial for cancer diagnosis and therapy. However, methods for detecting hypoxia are limited, often relying on single-detection systems. In this study, we developed a dual-lock-based fluorescent probe that selectively exhibits strong green fluorescence under hypoxic conditions due to simultaneous activity of nitroreductases (NTRs) and hydrogen sulfide (H2S), with a high signal-to-background ratio. The biocompatibility and photophysical properties of the probes were thoroughly investigated through both extracellular and intracellular experimental analyses. Among the synthesized naphthalimide-based probes, the dual-detection probe DNNC demonstrated excellent selectivity and sensitivity to the simultaneous activity of NTR/H2S compared to other single-detection probes. The performance of DNNC was applied to various organ-derived cancer cells and tumor tissue models such as HeLa cell sparoids, enabling spatiotemporal confocal fluorescence imaging and quantitative analysis of hypoxic levels in cancer. Our development of DNNC is expected to significantly advance cancer diagnosis and treatment by molecularly detecting hypoxia associated with cancer aggressiveness, therapy resistance, and unfavorable prognosis.

Verification of the accuracy of Sentinel-1 for DEM extraction error analysis under complex terrain conditions

The successful launch of the Sentinel-1 satellite in 2014 brought a large amount of free SAR images to researchers and scholars, and its application in the fields of ocean monitoring, land use change, natural disaster monitoring and emergency response is becoming increasingly mature and precise. The main applications of InSAR can be categorized into surface deformation monitoring and DEM generation. Sentinel-1 was initially designed for surface deformation monitoring; thus, there are fewer relevant studies on the use of Sentinel-1 data for DEM extraction. However, as the only SAR satellite whose data are currently free and openly available and whose data are constantly updated, it is highly important to study its sources of error in the DEM generation process and the accuracy of its products. In addition, the SAR data provided by the Sentinel-1 satellite has the advantages of high resolution, all-day, all-weather, providing a large data source for DEM production. Taking the Ankang area as an example, this paper analyzes the influence of the InSAR spatiotemporal baseline, ground cover, terrain factors, SAR imaging and other factors on the accuracy of the Sentinel-1-extracted DEM using multisource ground observation data to validate its feasibility for terrain mapping in complex terrain. Finally, we look forward to how to effectively improve the quality of Sentinel-1 DEM products to provide guidance and a reference for subsequent research on DEM extraction using Sentinel-1 SAR images and designation of Sentinel-1 C satellite’s parameters.

2D spatiotemporal passive cavitation imaging and evaluation during ultrasound thrombolysis based on diagnostic ultrasound platform

Acoustic cavitation plays a critical role in various biomedical applications. However, uncontrolled cavitation can lead to undesired damage to healthy tissues. Therefore, real-time monitoring and quantitative evaluation of cavitation dynamics is essential for understanding underlying mechanisms and optimizing ultrasound treatment efficiency and safety. The current research addressed the limitations of traditionally used cavitation detection methods by developing introduced an adaptive time-division multiplexing passive cavitation imaging (PCI) system integrated into a commercial diagnostic ultrasound platform. This new method combined real-time cavitation monitoring with B-mode imaging, allowing for simultaneous visualization of treatment progress and 2D quantitative evaluation of cavitation dosage within targeted area. An improved delay-and-sum (DAS) algorithm, optimized with a minimum variance (MV) beamformer, is utilized to minimize the side lobe effect and improve the axial resolution typically associated with PCI. In additional to visualize and quantitatively assess the cavitation activities generated under varied acoustic pressures and microbubble concentrations, this system was specifically applied to perform 2D cavitation evaluation for ultrasound thrombolysis mediated by different solutions, e.g., saline, nanodiamond (ND) and nitrogen-annealed nanodiamond (N-AND). This research aims to bridge the gap between laboratory-based research systems and real-time spatiotemporal cavitation evaluation demands in practical uses. Results indicate that this improved 2D cavitation monitoring and evaluation system could offer a useful tool for comprehensive evaluating cavitation-mediated effects (e.g., ultrasound thrombolysis), providing valuable insights into in-depth understanding of cavitation mechanisms and optimization of cavitation applications.

Attention-based CNN-BiLSTM for sleep state classification of spatiotemporal wide-field calcium imaging data

BackgroundWide-field calcium imaging (WFCI) with genetically encoded calcium indicators allows for spatiotemporal recordings of neuronal activity in mice. When applied to the study of sleep, WFCI data are manually scored into the sleep states of wakefulness, non-REM (NREM) and REM by use of adjunct EEG and EMG recordings. However, this process is time-consuming, invasive and often suffers from low inter- and intra-rater reliability. Therefore, an automated sleep state classification method that operates on spatiotemporal WFCI data is desired. New methodA hybrid network architecture consisting of a convolutional neural network (CNN) to extract spatial features of image frames and a bidirectional long short-term memory network (BiLSTM) with attention mechanism to identify temporal dependencies among different time points was proposed to classify WFCI data into states of wakefulness, NREM and REM sleep. ResultsSleep states were classified with an accuracy of 84 % and Cohen’s κ of 0.64. Gradient-weighted class activation maps revealed that the frontal region of the cortex carries more importance when classifying WFCI data into NREM sleep while posterior area contributes most to the identification of wakefulness. The attention scores indicated that the proposed network focuses on short- and long-range temporal dependency in a state-specific manner. Comparison with existing methodOn a held out, repeated 3-hour WFCI recording, the CNN-BiLSTM achieved a κ of 0.67, comparable to a κ of 0.65 corresponding to the human EEG/EMG-based scoring. ConclusionsThe CNN-BiLSTM effectively classifies sleep states from spatiotemporal WFCI data and will enable broader application of WFCI in sleep research.

Closed Bipolar Nanoelectrode Array for Ultra-Sensitive Detection of Alkaline Phosphatase and Two-Dimensional Imaging of Epidermal Growth Factor Receptors.

The combination of closed bipolar electrodes (cBPE) with electrochemiluminescence (ECL) imaging has demonstrated remarkable capabilities in the field of bioanalysis. Here, we established a cBPE-ECL platform for ultrasensitive detection of alkaline phosphatase (ALP) and two-dimensional imaging of epidermal growth factor receptor (EGFR). This cBPE-ECL system consists of a high-density gold nanowire array in anodic aluminum oxide (AAO) membrane as the cBPE coupled with ECL of highly luminescent cadmium selenide quantum dots (CdSe QDs) luminophores to achieve cathodic electro-optical conversion. When an enzyme-catalyzed amplification effect of ALP with 4-aminophenyl phosphate monosodium salt hydrate (p-APP) as the substrate and 4-aminophenol (p-AP) as the electroactive probe is introduced, a significant improvement of sensing sensitivity with a detection limit as low as 0.5 fM for ALP on the cBPE-ECL platform can be obtained. In addition, the cBPE-ECL sensing system can also be used to detect cancer cells with an impressive detection limit of 50 cells/mL by labeling ALP onto the EGFR protein on A431 human epidermal cancer cell membranes. Thus, two-dimensional (2D) imaging of the EGFR proteins on the cell surface can be achieved, demonstrating that the established cBPE-ECL sensing system is of high resolution for spatiotemporal cell imaging.

High Spatiotemporal Resolution STEM Imaging at High Temperature

High Spatiotemporal Resolution STEM Imaging at High Temperature

Image Sensing of Gaseous Acetone Using Secondary Alcohol Dehydrogenase-Immobilized Mesh for Exhaled Air.

Human-borne acetone is a potent marker of lipid metabolism. Here, an enzyme immobilization method for secondary alcohol dehydrogenase (S-ADH), which is suitable for highly sensitive and selective biosensing of acetone, was developed, and then its applicability was demonstrated for spatiotemporal imaging of concentration distribution. After various investigations, S-ADH-immobilized meshes could be prepared with less than 5% variation by cross-linking S-ADH with glutaraldehyde on a cotton mesh at 40 °C for 15 min. Furthermore, high activity was obtained by adjusting the concentration of the coenzyme nicotinamide adenine dinucleotide (NADH) solution added to the S-ADH-immobilized mesh to 500 μM and the solvent to a potassium phosphate buffer solution at pH 6.5. The gas imaging system using the S-ADH-immobilized mesh was able to image the decrease in NADH fluorescence (ex 340 nm, fl 490 nm) caused by the catalytic reaction of S-ADH and the acetone distribution in the concentration range of 0.1-10 ppm-v, including the breath concentration of healthy people at rest. The exhaled breath of two healthy subjects at 6 h of fasting was quantified as 377 and 673 ppb-v, which were consistent with the values quantified by gas chromatography-mass spectrometry.

CRISPR/Pepper-tDeg: A Live Imaging System Enables Non-Repetitive Genomic Locus Analysis with One Single-Guide RNA.

CRISPR-based genomic-imaging systems have been utilized for spatiotemporal imaging of the repetitive genomic loci in living cells, but they are still challenged by limited signal-to-noise ratio (SNR) at a non-repetitive genomic locus. Here, an efficient genomic-imaging system is proposed, termed CRISPR/Pepper-tDeg, by engineering the CRISPR sgRNA scaffolds with the degron-binding Pepper aptamers for binding fluorogenic proteins fused with Tat peptide derived degron domain (tDeg). The target-dependent stability switches of both sgRNA and fluorogenic protein allow this system to image repetitive telomeres sensitively with a 5-fold higher SNR than conventional CRISPR/MS2-MCP system using "always-on" fluorescent protein tag. Subsequently, CRISPR/Pepper-tDeg is applied to simultaneously label and track two different genomic loci, telomeres and centromeres, in living cells by combining two systems. Given a further improved SNR by the split fluorescent protein design, CRISPR/Pepper-tDeg system is extended to non-repetitive sequence imaging using only one sgRNA with two aptamer insertions. Neither complex sgRNA design nor difficult plasmid construction is required, greatly reducing the technical barriers to define spatiotemporal organization and dynamics of both repetitive and non-repetitive genomic loci in living cells, and thus demonstrating the large application potential of this genomic-imaging system in biological research, clinical diagnosis and therapy.

Duo-Chol: A Photoconvertible Live Cell Imaging Tool for Tracking Cholesterol.

Investigating cholesterol trafficking pathways continues to be of significant scientific interest owing to its homeostasis being associated with several debilitating cardiovascular and neurodegenerative diseases including atherosclerosis, Niemann-Pick's disease, Alzheimer's disease, and Parkinson's disease. To further our understanding of cholesterol trafficking, it is imperative to develop new fluorescent probes that possess improved photostability, low efflux, and high spatial and temporal resolution for live-cell imaging. In this study, we developed a photoconvertible fluorescent cholesterol analog, Duo-Chol, enabling the improved spatiotemporal fluorescence imaging of the dynamic localization of cholesterol in live cells. This tool provides a unique and powerful approach to interrogating cholesterol dynamics, addressing the limitations of existing methods, and expanding our ability to probe the biological role of sterols in living cells.

Myosin XI-mediated BIK1 recruitment to nanodomains facilitates FLS2–BIK1 complex formation during innate immunity in Arabidopsis

Plants rely on immune receptor complexes at the cell surface to perceive microbial molecules and transduce these signals into the cell to regulate immunity. Various immune receptors and associated proteins are often dynamically distributed in specific nanodomains on the plasma membrane (PM). However, the exact molecular mechanism and functional relevance of this nanodomain targeting in plant immunity regulation remain largely unknown. By utilizing high spatiotemporal resolution imaging and single-particle tracking analysis, we show that myosin XIK interacts with remorin to recruit and stabilize PM-associated kinase BOTRYTIS-INDUCED KINASE 1 (BIK1) within immune receptor FLAGELLIN SENSING 2 (FLS2)-containing nanodomains. This recruitment facilitates FLS2/BIK1 complex formation, leading to the full activation of BIK1-dependent defense responses upon ligand perception. Collectively, our findings provide compelling evidence that myosin XI functions as a molecular scaffold to enable a spatially confined complex assembly within nanodomains. This ensures the presence of a sufficient quantity of preformed immune receptor complex for efficient signaling transduction from the cell surface.

Enhancement of pH imaging of light addressable potentiometric sensor by colloidal spherical lens array

Visualizing the distribution of pH in a solution is an important basis for monitoring reaction processes and metabolic changes in micro-biochemical species. Light addressable potentiometric sensor (LAPS), as a simple structure, flexible imaging, low-cost and label-free electrochemical sensor, has been widely used to pH imaging. However, its spatiotemporal resolution is greatly affected by the frequency of modulation light, thus limiting its imaging performance. In this study, LAPS device modified by well-ordered and low-cost polystyrene (PS) colloidal monolayer as spherical lens array is proposed to obtain higher available modulation frequency of the maximum photoresponse. The morphology, optical properties and the electric field distribution of PS colloidal spherical lens array were tested and analyzed. pH sensing, imaging of uniform pH solution, the spatiotemporal pH imaging and dynamic imaging of enzymatic reactions of LAPS modified by PS colloidal spherical lens array were tested and evaluated. The experimental results show that LAPS modified by PS colloidal spherical lens array not only has acceptable pH sensing, but also widens the modulation frequency of the maximum photoresponse to 40 kHz. LAPS modified by colloidal spherical lens array modulated at the frequency of 40 kHz shows acceptable sensitivity and linearity, good reproducibility and stability, high SNR and high-spatiotemporal pH imaging capability. This work can be suitable for high-spatiotemporal monitoring of the changes in electrolyte environmental pH caused by the reaction and metabolism of the biochemical species.

Catalytic hairpin assembly-based AIEgen/graphene oxide nanocomposite for fluorescence-enhanced and high-precision spatiotemporal imaging of microRNA in living cells

The low abundance, heterogeneous expression, and temporal changes of miRNA in different cellular locations pose significant challenges for both the detection sensitivity of miRNA liquid biopsy and intracellular imaging. In this work, we report an intelligently assembled biosensor based on catalytic hairpin assembly (CHA) and aggregation-induced emission (AIE), named as catalytic hairpin aggregation-induced emission (CHAIE), for the ultrasensitive detection and intracellular imaging of miRNA-155. To achieve such goal, tetraphenylethylene-N3 (TPE-N3) is used as AIE luminogen (AIEgen), while graphene oxide is introduced to quench the fluorescence. When the target miRNA is present, CHA reaction is triggered, causing the AIEgen to self-assemble with the hairpin DNA. This will restrict the intramolecular rotation of the AIEgen and produce a strong AIE fluorescence. Interestingly, CHAIE does not require any enzyme or expensive thermal cycling equipment, and therefore provides a rapid detection. Under optimal conditions, the proposed biosensor can determine miRNA in the concentration range from 2 pM to 200 nM within 30 min, with the detection limit of 0.42 pM. The proposed CHAIE biosensor in this work offers a low background signal and high sensitivity, making it applicable for highly precise spatiotemporal imaging of target miRNA in living cells.